Disulfide-Stabilized Multivalent Antibody Fusion Proteins
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Solution Overview
Problem
Current methods for producing dual specificity antibody fusion proteins face challenges such as increased clearance rates due to the lack of an Fc domain, steric hindrance issues, and inefficient production techniques like chemical cross-linking and complex protein engineering, which affect their in vivo half-life and stability.
Innovation Solution
A recombinantly produced multivalent antibody fusion protein comprising a Fab or Fab' fragment with a first specificity and two single domain antibodies (dAbs) linked by a disulfide bond, where the dAbs are connected to the Fab or Fab' via genetic fusion, providing extended half-life by binding to serum carrier proteins like human serum albumin.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of operation
If antibody fragments (Fab, Fv) are used instead of whole antibodies, then tissue penetration and pharmacokinetic properties are improved, but clearance rate increases due to lack of Fc domain
Solution Approach 1:
The patent merges the Fc domain with the Fab fragment through genetic fusion, creating a chimeric molecule that combines the advantages of both components. The Fc domain provides extended serum half-life while the Fab fragment maintains tissue penetration capabilities, resolving the contradiction between rapid clearance and prolonged circulation.
Solution Approach 2:
The invention creates a composite antibody molecule combining different functional domains (Fc and Fab) with distinct properties. This composite structure allows the molecule to simultaneously achieve both rapid tissue penetration (from Fab) and extended serum half-life (from Fc domain), addressing the contradiction between these two characteristics.
2Adaptability or versatility
If chemical cross-linking is used to produce bispecific antibodies, then dual specificity is achieved, but production yield is poor and chromatographic separation is required
Solution Approach 1:
The patent replaces chemical cross-linking methods with genetic fusion techniques. Instead of using chemical agents to cross-link separate antibody components, the invention uses genetic engineering to fuse multiple specificities into a single continuous protein sequence, eliminating the need for chemical cross-linking agents and subsequent chromatographic separation steps.
Solution Approach 2:
The invention extracts and eliminates the need for chemical cross-linking agents and complex purification procedures by using direct genetic fusion. This approach removes the harmful and inefficient chemical cross-linking step, allowing for straightforward recombinant protein production with high yields.
3Adaptability or versatility
If protein engineering techniques (knobs-into-holes) are used to make bispecific antibodies, then dual specificity is achieved, but the approach becomes highly elaborate
Solution Approach 1:
The patent extracts and eliminates complex protein engineering techniques such as knobs-into-holes engineering. By using direct genetic fusion, the invention removes the need for elaborate structural modifications and complex engineering procedures, simplifying the overall process while maintaining dual specificity.
Solution Approach 2:
The invention segments the antibody molecule into distinct functional domains (Fc and Fab) that can be independently designed and then fused together. This segmentation allows for modular construction of bispecific antibodies, reducing the overall engineering complexity compared to attempting to create entirely new protein structures.
4Ease of manufacture
If diabodies or scFv are used for bispecific antibodies, then production is simplified, but stability characteristics are inappropriate
Solution Approach 1:
The patent creates a composite structure combining the simplicity of fusion protein production with the stability of properly folded antibody domains. By ensuring correct domain arrangement and folding in the chimeric molecule, the invention achieves both ease of manufacture and appropriate stability characteristics.
Solution Approach 2:
The invention changes the structural parameters of the antibody molecule by properly arranging Fc and Fab domains in a stable configuration. This structural reparameterization ensures molecular stability while maintaining the production simplicity of fusion protein techniques.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The dual specificity antibody fusion proteins achieve improved stability and extended in vivo half-life by effectively binding to serum carrier proteins, enhancing their therapeutic potential and diagnostic capabilities.
Implementation Method 1
two single domain antibodies (dAbs) which are a VH/VL pair with specificity for a second antigen of interest, wherein the two single domain antibodies are linked by a disulfide bond between two cysteine residues, one in VH and one in VL
Implementation Method 2
A recombinantly produced multivalent antibody fusion protein comprising a Fab or Fab' fragment with a first specificity and two single domain antibodies (dAbs) linked by a disulfide bond, where the dAbs are connected to the Fab or Fab' via genetic fusion, providing extended half-life by binding to serum carrier proteins like human serum albumin.
Data Source
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Figure 2A~2B
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AI summary
A multivalent antibody fusion protein which comprises an immunoglobulin moiety, for example a Fab or Fab' fragment, with a first specificity for an antigen of interest, and further comprises two single domain antibodies (dAb) with specificity for a second antigen of interest which are a VH/VL pair, wherein the two single domain antibodies are linked by a disulfide bond. Also provided are particular dual specificity antibody fusion proteins and other antibody fragments which are stabilised by a disulfide bond.